TUESDAY June 4th - pick up packages in Fisher storeroom in WELCH. Sell on the black market and pocket the CA$H.
Pick up Primers at the MBB front desk - again (060313 - Monday)
Print out copies of the Nanodrop, DNA Sequencing & Primer Dilution protocols - 2 pages per sheet so that they can go in the notebooks.
Get the autoclaved water and aliquot several into 1.7 ml centrifuge tube, label and store in -20degC. We will have the students use it today (Monday) for primer dilution.
When doing Nanodrop and SUbmit to DNA ssequencing of (pGBR22, pGFP and pmCHerry) - the students should be able to track back exactly which tube they used.
Thursday May 30th @ 1-5 pm
PURCHASING at BioScience Storeroom (NHB) - for Thurs -
Gold watches, bling, etc.
Porsche 911, 1962 Ferrari GTO, etc.
VDS staff
1
25 nmoles
IDT
via ICMB
VDS account
$3.57
pLIC-for
TGTGAGCGGATAACAATTCC
VDS staff
1
25 nmoles
IDT
via ICMB
VDS account
$3.39
pLIC-rev
AGCAGCCAACTCAGCTTCC
VDS staff
1
Biosci Storeroom
$37.51
Tryptone
VDS staff
1
c0180
?
Bio Sci Storeroom
$10.21
Sodium Hydroxide Pellets
VDS staff
1
kit
Sigma
Biosci Storeroom
$61.80
PCR Clean up Kit, Genelute
EP008
VDS staff
1
kit
Sigma
Biosci Storeroom
$337.58
Plasmid Mini Prep 350 isolations
VDS staff
1
Qiagen
Biosci Storeroom
$338.68
HiSpeed Plasmid Midi
VDS staff
1
10,000 U
NEB
Biosci Storeroom
$49.68
EcoRI HF
E0011
VDS staff
1
PK
N0446S
NEB
Biosci Storeroom
$138.00
dNTPs - individuals (Nucleotide Solution Pack)
E0065
VDS staff
1
PK
N0447S
NEB
Biosci Storeroom
$49.68
dNTPs - mix for Taq PCR 8 μmol of each
E0066
VDS staff
1
400 U
M0267S
NEB
Biosci Storeroom
$54.28
standard Taq DNA polymerase with ThermoPol
E0041
VDS staff
1
150 U
NEB
Biosci Storeroom
$57.04
T4 DNA polymerase
VDS staff
1
500 ug
E0060
NEB
Biosci Storeroom
$184.00
1 kb ladder
VDS staff
6
Biosci Storeroom
$0.54
Marker, Sharpie, Ultra Fine Point, Black
VDS staff
?? Needed
1
kit (70)
Sigma
Biosci Storeroom
$101.66
GenElute gel Extraction (70)
spray bottles - 2
WET LAB WORK
Prep collirollers - split into 2 bottles and rinse then autoclave
Remove ice from Top Loading freezer. Remove ice on top and on the bottom - maybe de-thaw the whole thing.
Organize and re-label freezer boxes for us (ideally we want each 'type' of reagent to have its own box. We can use up to 3 metal racks. Use new plastic boxes when needed.
New Boxes to Make for Top Loading freezer: Old Primers 2, Modifying Enzymes, Buffers 2
re-locate all of the buffers to the metal racks.
Make a new VDS Class Plasmids box for the upright -20degC
Organize dry chemicals above weighing bench
Prep DNA ladders - 100bp and 1 kb (see protocol on GDocs). Then aliquot into 5-10 different tubes, well labeled
On Wikispaces page, Update the 1 month vs 2 month people - move 1 month people to separate list -
Assign students to Red, Green or Purple for Plasmid work and Transformation Efficiency
Update Laptops and Desktops
Clean off old icons from Desktops of Desktop computers
Do Windows Updates
Start up laptops and do updates on them. Remove security credentials.
Install both printers on each laptop and each Desktop (see below)
Organize and print new protocols (updated) for the white protocols binder
- in the back of the lab (on top of fire cabinet)
- use plastic sheet protectors
Clean water baths (the big ones - probably need 2 people for this). Only use Alconox soap - no bleach or anything harsh.
Get gel extra dryer from Malcolm Brown, Jr. Lab. Test functionality.
Consolidate Kits - Miniprep, Midiprep, PCR Cleanup. Put spare solutions into 1 box and label it spare solutions for that Type of Kit. Verify that all components are present (or in sufficient number
Count protein gel spacer plates vs. short plates vs. combs.
Check on how much glassware we have. If low - purchase more.
send pGBR22 samples to DNA Sequencing so that we can figure out which ones had the contamination (maybe have the students do this)
FOR LATER:
Dispense collirollers into individual 600 ul tubes with each containing ~6 beads. Do this sterily (student to do?)
Check lab safety completion for all students
-- see the printed sheets in the lab binder and then cross check them to the Wikispaces 'Required Lab Safety' page
- cross reference to UTEIDs of Class Roster (can be found in VDSstaff@gmail.com account
Check Ice Room access list -
Printer Installation
Login as Mentor, Start Menu,
Devices And printers
Add New Printer
Add "local" printer
Create New Port --> standard TCP pathway
Enter IP 128.82.195.186
Printer Name Lexmark C543
Add Printer (if can't find printer do Window Update)
For installation of a different Printer (such as Dell) change the IP address and Printer Name
Printer Name: Lexmark C543 IP: 128.83.195.186
Printer Name: Dell IP: 129.116.152.49
SOLUTION RECIPES
Make LB - 2Liters - and autoclave
Make 5 M NaCl - 500 ml in water, sterile filter into a bottle, label, store at Room Temp
Make Kanamycin- stocks 50ug/ul store in -20degC, Make 30 tubes of 500 ul each. Sterile filter with syringe filterbefore aliquoting into tubes. Label. Store in -20degC
Make IPTG - 1 M IPTG stock to make. Make 30 tubes of this with 500 ul in each. Label. Store in -20degC
Make Lysozyme - 20 aliquots of 500 ul at 50 mg/ml in 1.7 ml tubes. Label. Store in -20degC
Make TCEP - make ~50 x 100ul aliquots of 500 mM aliquots medium size centrifuge tubes. Label, put in a box and Store at Room Temp in cubby.
Make PMSF - 30 x 500 ul aliquots of 10mM in Isopropanol 1.7 ml tubes. Label. store in -20degC
Make Imidazole - 1M stock make in beaker of 500 ml of water, then bottle top sterile filter it into a bottle, store at Room Temp
Lysis Buffer for Sonication (make 500 ml)
(see GDocs 'Solutions&Dilutions' in /Misc folder for calculations)
Tris 100 mM
NaCl 300 mM
Imidazole 30 mM
pH 8.0 w/ NaOH
label, store at 4degC
NEW Wash and Elution Buffers and Storage Buffer(see GDocs 'Solutions&Dilutions' in /Misc folder for calculations)
Wash Buffer
Tris 100 mM
NaCl 300 mM
Imidazole 30 mM
pH 8.0 w/ NaOH
Elution Buffer
Tris 100 mM
NaCl 300 mM
Imidazole 300 mM
pH 8.0 w/ NaOH
FPLC column buffer - tris Also 'storage' buffer
Tris 50 mM
NaCl 150 mM
pH 8.0 w/ NaOH
Then filter sterilize
Then degass**
6X SDS Page Buffer from 5 Prime Protocol
30 ml of glycerol – add first so that you can see the volume (viscous)
(In a separate tube) - 30 ml of Nanopure water
--- 1.635 g Tris powder
--- 3 g SDS poweder
--- 0.3 g Bromophenol blue
----2.3136 g DTT
Mix contents of second tube into tube with glycerol
Add some more Nanopure water to 50 ml total final volume
DANIEL, MICHAEL, ANDREW, RUIFEI
VDS Staff Meeting:
TUESDAY June 4th - pick up packages in Fisher storeroom in WELCH. Sell on the black market and pocket the CA$H.
Pick up Primers at the MBB front desk - again (060313 - Monday)
Print out copies of the Nanodrop, DNA Sequencing & Primer Dilution protocols - 2 pages per sheet so that they can go in the notebooks.
Get the autoclaved water and aliquot several into 1.7 ml centrifuge tube, label and store in -20degC. We will have the students use it today (Monday) for primer dilution.
When doing Nanodrop and SUbmit to DNA ssequencing of (pGBR22, pGFP and pmCHerry) - the students should be able to track back exactly which tube they used.
Thursday May 30th @ 1-5 pm
PURCHASING at BioScience Storeroom (NHB) - for Thurs -
Gold watches, bling, etc.Porsche 911, 1962 Ferrari GTO, etc.
WET LAB WORK
Prep collirollers - split into 2 bottles and rinse then autoclaveRemove ice from Top Loading freezer. Remove ice on top and on the bottom - maybe de-thaw the whole thing.
Organize and re-label freezer boxes for us (ideally we want each 'type' of reagent to have its own box. We can use up to 3 metal racks. Use new plastic boxes when needed.
New Boxes to Make for Top Loading freezer: Old Primers 2, Modifying Enzymes, Buffers 2
re-locate all of the buffers to the metal racks.
Make a new VDS Class Plasmids box for the upright -20degC
Organize dry chemicals above weighing bench
Prep DNA ladders - 100bp and 1 kb (see protocol on GDocs). Then aliquot into 5-10 different tubes, well labeled
On Wikispaces page, Update the 1 month vs 2 month people - move 1 month people to separate list -
Update Laptops and Desktops
Organize and print new protocols (updated) for the white protocols binder
- in the back of the lab (on top of fire cabinet)
- use plastic sheet protectors
Clean water baths (the big ones - probably need 2 people for this). Only use Alconox soap - no bleach or anything harsh.
Get gel extra dryer from Malcolm Brown, Jr. Lab. Test functionality.
Consolidate Kits - Miniprep, Midiprep, PCR Cleanup. Put spare solutions into 1 box and label it spare solutions for that Type of Kit. Verify that all components are present (or in sufficient number
Count protein gel spacer plates vs. short plates vs. combs.
Check on how much glassware we have. If low - purchase more.
send pGBR22 samples to DNA Sequencing so that we can figure out which ones had the contamination (maybe have the students do this)
FOR LATER:
Dispense collirollers into individual 600 ul tubes with each containing ~6 beads. Do this sterily (student to do?)Check lab safety completion for all students
-- see the printed sheets in the lab binder and then cross check them to the Wikispaces 'Required Lab Safety' page- cross reference to UTEIDs of Class Roster (can be found in VDSstaff@gmail.com account
Check Ice Room access list -
Printer Installation
For installation of a different Printer (such as Dell) change the IP address and Printer Name
Printer Name: Lexmark C543 IP: 128.83.195.186
Printer Name: Dell IP: 129.116.152.49
SOLUTION RECIPES
Make LB - 2Liters - and autoclave
Make 5 M NaCl - 500 ml in water, sterile filter into a bottle, label, store at Room Temp
Make Kanamycin- stocks 50ug/ul store in -20degC, Make 30 tubes of 500 ul each. Sterile filter with syringe filterbefore aliquoting into tubes. Label. Store in -20degC
Make IPTG - 1 M IPTG stock to make. Make 30 tubes of this with 500 ul in each. Label. Store in -20degC
Make Lysozyme - 20 aliquots of 500 ul at 50 mg/ml in 1.7 ml tubes. Label. Store in -20degC
Make TCEP - make ~50 x 100ul aliquots of 500 mM aliquots medium size centrifuge tubes. Label, put in a box and Store at Room Temp in cubby.
Make PMSF - 30 x 500 ul aliquots of 10mM in Isopropanol 1.7 ml tubes. Label. store in -20degC
Make Imidazole - 1M stock make in beaker of 500 ml of water, then bottle top sterile filter it into a bottle, store at Room Temp
Lysis Buffer for Sonication (make 500 ml)
(see GDocs 'Solutions&Dilutions' in /Misc folder for calculations)
Tris 100 mM
NaCl 300 mM
Imidazole 30 mM
pH 8.0 w/ NaOH
label, store at 4degC
NEW Wash and Elution Buffers and Storage Buffer(see GDocs 'Solutions&Dilutions' in /Misc folder for calculations)
Wash Buffer
Tris 100 mM
NaCl 300 mM
Imidazole 30 mM
pH 8.0 w/ NaOH
Elution Buffer
Tris 100 mM
NaCl 300 mM
Imidazole 300 mM
pH 8.0 w/ NaOH
FPLC column buffer - tris Also 'storage' buffer
Tris 50 mM
NaCl 150 mM
pH 8.0 w/ NaOH
Then filter sterilize
Then degass**
6X SDS Page Buffer from 5 Prime Protocol
30 ml of glycerol – add first so that you can see the volume (viscous)
(In a separate tube) - 30 ml of Nanopure water
--- 1.635 g Tris powder
--- 3 g SDS poweder
--- 0.3 g Bromophenol blue
----2.3136 g DTT
Mix contents of second tube into tube with glycerol
Add some more Nanopure water to 50 ml total final volume
Mentor Notes from Spring 13
Mentors
|| 1 || 25 nmoles || || IDT || via ICMB || VDS account || $3.57 || $3.57 || pLIC-for || |||||| TGTGAGCGGATAACAATTCC || ||